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. 2011 Jan 13;286(11):9598–9611. doi: 10.1074/jbc.M110.195909

FIGURE 7.

FIGURE 7.

Effect of proteasomal inhibition by MG132 and lactacystin on expression of YFP-tagged WT and W391A-CaV2.2 in SCG somata and neurites, using CFP-CaV2.2 as an internal control. A, examples of SCG neuron somata expressing CFP-CaV2.2(WT) (left), together with YFP-CaV2.2(W391A) (right), injected after 6 h in culture, and imaged 18 h later, in the presence of 50 nm (top), 500 nm (middle), and 1 μm (bottom) MG132. Scale bars, 20 μm. Note that the image plane does not go through the nucleus in all cases. B, bar chart of the ratio of YFP/CFP fluorescence in cell bodies, from data such as those in A, for YFP-CaV2.2(WT) (black bar; n = 14), YFP-CaV2.2(W391A) (white bar; n = 12), and YFP-CaV2.2(W391A) together with 50 nm (light gray bar; n = 13), 250 nm (dark gray bar; n = 13), or 1 μm (hatched bar; n = 13) MG132. All experiments also included CFP-CaV2.2(WT). The statistical significance between YFP-CaV2.2(W391A) in the absence and presence of MG132 is shown: * p < 0.05, one-way analysis of variance and Bonferroni's post-test. C, bar chart of the ratio of YFP/CFP fluorescence in neurites, for YFP-CaV2.2(WT) (black bar; n = 17), YFP-CaV2.2(W391A) (white bar; n = 17), and YFP-CaV2.2(W391A) together with 50 nm (light gray bar; n = 13), 250 nm (dark gray bar; n = 14), or 1 μm (hatched bar; n = 19) MG132. All experiments also included CFP-CaV2.2(WT). The statistical significances are shown as follows: *, p < 0.05; **, p < 0.01; ***, p < 0.001, one-way ANOVA and Bonferroni's post-test. D, examples of SCG neuron somata expressing CFP-CaV2.2(WT) (left), together with YFP-CaV2.2(W391A) or YFP-CaV2.2(WT) (right), injected after 6 h in culture, and imaged 18 h later, in the presence of DMSO or lactacystin (10 μm), as indicated. Scale bars, 20 μm. Note that the image plane goes through the nucleus in all cases. E, bar chart of the ratio of YFP/CFP fluorescence in cell bodies, from data such as those in D, for YFP-CaV2.2(WT) + DMSO (black bar; n = 8), CaV2.2(WT) + lactacystin (white bar; n = 7), YFP-CaV2.2(W391A) + DMSO (light gray bar; n = 11), and CaV2.2(W391A) + lactacystin (dark gray bar; n = 11). All experiments also included CFP-CaV2.2(WT). The statistical significances are shown: ***, p < 0.001; **, p < 0.01, one-way analysis of variance and Bonferroni's post-test. Error bars, S.E.